Untitled

Gospodinov, G. G.; Stancheva, M. G.

doi:10.1023/a:1025118532216

Cited by 5 publications

(2 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A protocol for the cell-free DNA repair system developed in our laboratory was used [19]. Cell-free reactions contained 300 ng UV irradiated pBlueScript (ampicillin resistant) repair substrate, 300 ng unirradiated pEGFP (kanamycin resistant) internal control, 45 mM HEPES-KOH (pH 7.8), 70 mM KCl, 7.4 mM MgCl 2 , 0.9 mM dithiothreitol, 0.4 mM EDTA, 2 mM ATP, 25 lM each of dGTP, dATP, dTTP and dCTP, 40 mM phosphocreatine, 2.5 lg creatine phosphokinase, 18 lg bovine serum albumin, and 100 lg protein extract in a final volume of 50 ll.…”

Section: Cell-free Repair Assaymentioning

confidence: 99%

“…For these experiments, we chose to use K562 cells because they show similar to HeLa cell cycle distribution (54% in G1 and 29% in S phase), but are easily grown in the large quantity needed to prepare the extracts. The extent of repair was determined by the restoration of the colony forming ability of the damaged plasmids after transformation in E. coli [19]. Plasmids carrying UV lesions cannot replicate after transformation of E. coli in the absence of SOS induction and for this reason do not support colony formation on selective medium [20,21].…”

Section: Protein Extracts Of Cells Synchronized In S Phase Reproducedmentioning

confidence: 99%

See 1 more Smart Citation

Cells synchronized in S phase show increased rate of repair of UV damaged plasmids

Gospodinov

Anachkova

2004

FEBS Letters

View full text Add to dashboard Cite

The capacity for nucleotide excision repair of cells synchronized in S phase and unsynchronized cells was compared by the host cell reactivation assay and the cell-free repair system. HeLa cells were transfected with in vitro damaged by UV irradiation pEGFP and the repair capacity was determined by the number of fluorescent cells. In the cell-free repair system, the repair capacity of protein extracts isolated from K562 cells was determined by measuring the transformation efficiency of UV irradiated pBlueScript incubated in the extracts. In both cases, the repair capacity of the cells synchronized in S phase cells was 30-50% higher than the repair capacity of unsynchronized cells.

show abstract

Section: Cell-free Repair Assaymentioning

confidence: 99%

Section: Protein Extracts Of Cells Synchronized In S Phase Reproducedmentioning

confidence: 99%